Method of treating polyester fabrics

ABSTRACT

This invention relates to a method of reducing the pilling propensity or color clarity of polyester containing fabrics and/or garments, which method comprises treating the fabric with a polyester hydrolytic enzyme without the presence of a detergent.

TECHNICAL FIELD

This invention relates to a method of reducing the pilling propensityand/or improves the color clarity of polyester fabrics and/or garments,which method comprises treating the fabric with a polyester hydrolyticenzyme without the presence of a detergent. The invention also relatesto a method of bio-polishing polyester containing fabrics and garments.

BACKGROUND ART

Poly(ethylene terephthalate) fibers accounts for the main part of thepolyester applied by the textile industry. The fibers are produced by,e.g., poly-condensation of terephthalic acid and ethylene glycol, anddrawing of fibers from a melt.

Because of its strength, polyester fabrics and/or garments are subjectto pill formation, and possibly the most important of thecloth-finishing processes applied to polyester staple-fibre materialsare those designed for control of pilling. All staple-fibre materialstend to form small balls or “pills” of entangled fibres at the clothsurface, when subjected to mild abrasion during wash and wear. If thefabric contains a substantial proportion of fibres having highresistance to flexural abrasion, the pills may be retained on thesurface of the cloth in sufficient numbers to produce an unpleasanthandle and appearance.

According to Hatch in Textile Science (St. Paul: West PublishingCompany, 1993. pages 52-53, 218, 420) the mechanism of pilling is asfollows: i) Mechanical action causes fibers to migrate out of the fabricbody to the surface, ii) Further action causes the surface fibers torotate around other protruding fibers forming pills, iii) Additionalaction may continue to form more pills or to sever fibers anchoringpills. The pilling propensity of the fabric depends on the surface fuzzformation, the rate of fuzz entanglement, and finally the rate of pillsbreaking off. The rate of the pills breaking off is directly related tothe tenacity of the anchor fibers.

Bio-polishing is a finishing process where a textile fiber or yarn istreated with an enzyme to impart properties such as anti-pilling,softness and smoothness. This concept was initially developed in Japanwhere the first experiments were performed on cotton woven fabrics usingcellulases.

Pilling prevention is an ongoing challenge for manufacturers of cotton,polyester and blended fabrics. There is no simple solution to theproblem of pilling. In the textile industry, polyester fibers areproduced as medium- and high-tenacity filament yarns and as staplefibers of various lengths and fiber color to suit the kind of spinningmachinery found in the textile trade. Staple fibers are usually drawn togive medium tenacities, but may be spun from polymers of lower averagemolecular weight to give improved “pilling” performance at the expenseof some loss in abrasion resistance. Also, the finisher may reduce thepilling propensity of a fabric by the removal of protruding hairs fromthe surface of the cloth and by heat treatment to reduce the tendency ofthe fibres to migrate within the yarns.

Bazin, J. and Sasserod, S., 58ème Congrèss de l'Association desChimistes de l'lndustrie Textile Science, Mullhouse, France. Oct. 25,1991, discloses that a reduction in fuzz level which resulted in adramatic reduction in pilling on both 100% cotton and polyester cottonblends could be obtained using cellulase.

U.S. Pat. No. 5,997,584 discloses a method of reducing the pillingpropensity of polyester fabrics and/or garments, which method comprisestreating the fabric or garment with a terephtalic acid diethyl esterhydrolytic enzyme and/or an ethyleneglycol dibenzyl ester hydrolyticenzyme, and which method is carried out in presence of a detergent.

SUMMARY OF THE INVENTION

Pilling of fabric changes the aesthetic properties of textiles. Thesmoothness, color and general hand of fabric can be compromised.Therefore, it is an object of the present invention to provide animproved method of reducing the pilling propensity of polyester fabricsand garments. In particular, the invention provides that a group ofethyleneglycol dibenzyl ester (BEB) and/or terephtalic acid diethylester (ETE) hydrolytic enzymes are capable of reducing pillingpropensity of polyester fabrics and garments without the presence of adetergent.

The inventors have found that enzymatic bio-polishing offers a durablefinish for pill prevention. It was demonstrated (see Example 3) thatethyleneglycol dibenzyl ester (BEB) and/or terephtalic acid diethylester (ETE) hydrolytic enzymes are capable of reducing pillingpropensity of polyester fabrics and garments without the presence of adetergent. More specifically, the inventors have demonstrated thatethyleneglycol dibenzyl ester (BEB) and/or terephtalic acid diethylester (ETE) hydrolytic enzymes within the group of esterases, preferablycutinases, are capable of reducing pilling propensity of polyesterfabrics and garments without the presence of a detergent. Also thecombination of ethyleneglycol dibenzyl ester (BEB) and/or terephtalicacid diethyl ester (ETE) hydrolytic enzymes was found to provideimproved bio-polishing/pilling of polyester and cotton blended fabrics.

Furthermore, the treatment with ethyleneglycol dibenzyl ester (BEB)and/or terephtalic acid diethyl ester (ETE) hydrolytic enzymes iscapable of color clarification of polyester fabrics and/or garmentswithout the presence of a detergent.

From experiments with use of an ETE hydrolytic enzyme and/or a BEBhydrolytic enzyme for reducing the pilling propensity, it has been foundthat the method of the present invention allow for improvements in colorclarification. Therefore, in a preferred embodiment, the method of theinvention may be carried out simultaneously with conventional colorclarification processes. Color clarification processes have beendescribed in, e.g., EP 220,016; WO 91/17243; WO 89/09259; WO 91/19807;WO 94/07998 and WO 96/29397.

The term “color clarification”, as used herein, refers to preservationof the initial colors throughout multiple washing cycles by removingfuzz and pills from the surface of garment and/or fabric. The colorclarification ability may be determined by measuring the reflectance ofthe textile.

Also, the method of the invention allows for improved soil releaseproperties, in particular of oily stains, probably due to increasedhydrophilicity of the polyester fibers.

Finally, the method of the invention was found to improve the antistaticproperties of polyester fabrics and/or garments.

Accordingly, in its first aspect, the invention provides a method ofreducing the pilling propensity of polyester fabrics and/or garments,which method comprises treating the fabric or garment with a terephtalicacid diethyl ester hydrolytic enzyme and/or an ethyleneglycol dibenzylester hydrolytic enzyme, and which method is carried out without thepresence of a detergent.

In a second aspect, the invention provides a method of colorclarification of polyester fabrics and/or garments, which methodcomprises treating the fabric or garment with a terephtalic acid diethylester hydrolytic enzyme and/or an ethyleneglycol dibenzyl esterhydrolytic enzyme, and which method is carried out without the presenceof a detergent.

In a third aspect, the inventions provides a method of bio-polishing apolyester containing fabric or garment, which method comprises treatingthe fabric or garment with an enzyme selected from the group consistingof a terephtalic acid diethyl ester hydrolytic enzyme (ETE hydrolyticenzyme), an ethyleneglycol dibenzyl ester hydrolytic enzyme (BEBhydrolytic enzyme), and combinations of the foregoing, wherein saidmethod is carried out without the presence of a detergent.

Other aspects of the invention will become apparent from the followingdetailed description and the claims.

BRIEF DESCRIPTION OF THE DRAWING

FIG. 1 shows the effect of dosage on weight loss for esterasedegradation of 100% polyester fabric. Conditions: 2 hoursLaunder-O-Meter treatment at 70° C., pH 8.

FIG. 2 shows the effect of dosage on pilling note at 2000 revolutionsfor 100% polyester fabric. Conditions: 2 hours Launder-O-Meter treatmentat 70° C., pH 8.

FIG. 3 shows the effect of dosage on HPLC area count of polyesterdegradation peaks for 100% polyester fabric. Conditions: 2 hoursLaunder-O-Meter treatment at 70° C., pH 8.

DETAILED DISCLOSURE OF THE INVENTION

The present invention is directed to a method of reducing the pillingpropensity of polyester fabrics and/or garments. The inventionfurthermore provides a method of improving the color clarity ofpolyester fabrics and/or garments. Further, the invention also relatesto a method of bio-polishing polyester and polyester containing fabricsor garments.

As used herein, the singular forms “a”, “an”, and “the” include pluralreferences unless the context clearly dictates otherwise. Thus, forexample, references to “terephtalic acid diethyl ester hydrolyticenzyme” include the use of one or more terephtalic acid diethyl esterhydrolytic enzymes.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which the invention belongs. Although any methods andmaterials similar or equivalent to those described herein can be used inthe practice or testing of the present invention, the preferred methodsand materials are now described. All publications mentioned herein areincorporated herein by reference for the purpose of disclosing anddescribing the material for which the reference was cited in connectionwith.

Polyester Fabrics or Garments

The polyester fabrics and/or garments treated according to the method ofthe invention may be any fabric or fabric blend comprising polyester,including microdenier polyester. Actually, the pilling propensity ismost pronounced in fabrics and/or garments comprising polyester fibersin blends with fibers of a different material. Polyester fabric blendsinclude at least in context of the invention polyester and cellulosicblends, such as polyester and cotton blends. Other important polyesterfabric blends (i.e., polyester containing blends) include polyester andwool blends, polyester and silk blends, polyester and acrylic blends,polyester and nylon blends, polyester and nylon and polyurethane blends,polyester and polyurethane blends (e.g., LYCRA™, SPANDEX™), rayon(viscose), cellulose acetate and tencel.

In a preferred embodiment the fabric is a fabric blend comprising morethan 50% (w/w) of polyester, in particular more than 75% (w/w) ofpolyester, more than 90% (w/w) of polyester, or more than 95% (w/w) ofpolyester. In a most preferred embodiment, the process of the inventionis applied to fabrics or garments consisting essentially ofpoly(ethylene terephthalate) polyester material, i.e., purepoly(ethylene terephthalate) polyester material.

Polyester Hydrolytic Enzymes

The method of the invention comprises treating the fabric or garmentwith a polyester hydrolytic enzyme. A certain group of enzymes arecapable of hydrolysing terephtalic acid diethyl ester (ETE) and/or anethyleneglycol dibenzyl ester (BEB), and therefore arepolyesterhydrolytic enzymes.

Determination of which enzymes are ETE and/or BEB hydrolytic enzyme maybe carried out as described in Example 1.

The method of the invention comprises treating the fabric or garmentwith an ETE hydrolytic enzyme and/or a BEB hydrolytic enzyme without thepresence of a detergent. In a preferred embodiment, the method of theinvention comprises treating the fabric or garment with an ETEhydrolytic enzyme. In another preferred embodiment, the method of theinvention comprises treating the fabric or garment with a BEB hydrolyticenzyme. The BEB hydrolytic enzyme may in particular be a BEB¹⁰hydrolytic enzyme or BEB³⁰ hydrolytic enzyme, as defined in Example 1,below. Preferably the ETE hydrolytic enzyme has a hydrolytic activity ofat least 50%, more preferably of at least 90% and most preferably of atleast 95%. Preferably the BEB¹⁰ or BEB³⁰ hydrolytic enzyme has ahydrolytic activity of at least 50%, more preferably at least 90% andmost preferably at least 95%. In a most preferred embodiment both theBEB¹⁰, BEB³⁰ and ETE hydrolytic activity are at least 50%, morepreferably at least 90% and most preferably at least 95%.

In an embodiment of the method of the invention the polyester orpolyester containing fabric or garment is treated with an amount of ETEhydrolytic enzyme and/or a BEB hydrolytic enzyme in the range from about0.0001 to 10 mg enzyme protein/ml treating liquor, preferably 0.0006 to1 mg enzyme protein/ml treating liquor, especially in the range from0.006-1.2 mg enzyme protein/ml treating liquor. ETE hydrolytic enzymeand/or a BEB hydrolytic enzyme may be derived from any convenient originsuch as from bacterial, fungal, yeast, mammalian or plant origin.

Preferably the ETE hydrolytic enzyme and/or BEB hydrolytic enzyme isderived from a microbial source. In a more preferred embodiment, the ETEhydrolytic enzyme and/or BEB hydrolytic enzyme is derived from a strainof Candida, in particular, Candida antarctica and Candida cylindracea(syn. Candida rugosA), a strain of Humicola, in particular, Humicolainsolens, e.g., Humicola insolens DSM 1800, a strain of Pseudomonas, inparticular Pseudomonas cepacia.

As disclosed above, the enzymes may be derived from any origin,including, bacterial, fungal, yeast, mammalian or plant origin. The term“derived” means in this context that the enzyme may have been isolatedfrom an organism where it is present natively, i.e. the identity of theamino acid sequence of the enzyme are identical to a native enzyme. Theterm “derived” also means that the enzymes may have been producedrecombinantly in a host organism, the recombinant produced enzyme havingeither an identity identical to a native enzyme or having a modifiedamino acid sequence, e.g., having one or more amino acids which aredeleted, inserted and/or substituted, i.e., a recombinantly producedenzyme which is a mutant and/or a fragment of a native amino acidsequence or an enzyme produced by nucleic acid shuffling processes knownin the art. Within the meaning of a native enzyme are included naturalvariants. Furthermore, the term “derived” includes enzymes producedsynthetically by, e.g., peptide synthesis. The term “derived” alsoencompasses enzymes which have been modified, e.g., by glycosylation,phosphorylation, or by other chemical modification, whether in vivo orin vitro. The term encompasses an enzyme that has been isolated from anorganism where it is present natively, or one in which it has beenexpressed recombinantly in the same type of organism or another, orenzymes produced synthetically by, e.g., peptide synthesis. With respectto recombinantly produced enzymes the term “derived” refers to theidentity of the enzyme and not the identity of the host organism inwhich it is produced recombinantly.

The enzymes may also be purified. The term “purified” as used hereincovers enzymes free from other components from the organism from whichit is derived. The term “purified” also covers enzymes free fromcomponents from the native organism from which it is derived. Theenzymes may be purified, with only minor amounts of other proteins beingpresent. The expression “other proteins” relate in particular to otherenzymes. The term “purified” as used herein also refers to removal ofother components, particularly other proteins and most particularlyother enzymes present in the cell of origin of the enzyme of theinvention. The enzyme may be “substantially pure,” that is, free fromother components from the organism in which it is produced, that is, forexample, a host organism for recombinantly produced enzymes. Inpreferred embodiment, the enzymes are at least 75% (w/w) pure, morepreferably at least 80%, at least 85%, at least 90%, at least 95%, atleast 96%, at least 97%, at least 98%, or at least 99% pure. In anotherpreferred embodiment, the enzyme is 100% pure.

The enzyme may be in any form suited for the use in the treatmentprocess, such as, e.g., in the form of a dry powder or granulate, anon-dusting granulate, a liquid, a stabilized liquid, or a protectedenzyme. Granulates may be produced, e.g., as disclosed in U.S. Pat. No.4,106,991 and U.S. Pat. No. 4,661,452, and may optionally be coated bymethods known in the art. Liquid enzyme preparations may, for instance,be stabilized by adding stabilizers such as a sugar, a sugar alcohol oranother polyol, lactic acid or another organic acid according toestablished methods. Protected enzymes may be prepared according to themethod disclosed in EP 238,216.

The Treatment

The present invention provides a method of reducing the pillingpropensity of polyester fabrics and/or garments. Further, the presentinvention provides a method of improving the color clarification ofpolyester fabrics and/or garments. Furthermore, the invention alsorelates to a method of bio-polishing polyester and polyester blendfabrics or garments.

The treatment according to the present invention may be carried out atconditions chosen to suit the method according to principles well knownin the art. It will be understood that each of the reaction conditions,such as, e.g., concentration/dose of enzyme/substrate, pH, temperature,and time of treatment, may be varied, depending upon, e.g., the sourceof the enzyme, the type of substrate, the method in which the treatmentis performed.

The process of the invention may further comprise the addition of one ormore chemicals capable of improving the enzyme-substrate interaction (inorder to improve the substrate's accessibility and/or dissolve reactionproducts), which chemicals may be added prior to, or simultaneously withthe enzymatic treatment. Such chemicals may in particular be wettingagents and dispersing agents etc., or mixtures thereof.

Enzyme dosage must be a function of the enzyme(s) applied and thereaction time and conditions given. Preferably, the enzyme(s) may bedosed in a total amount of from about 0.05 micro grams per gram fabricsand/or garments to about 5000 micro grams per gram fabrics and/orgarments.

The enzymatic treatment may preferably be carried out in the temperaturerange of from about 30° C. to about 100° C., more preferably from about40° C. to about 90° C. The pH range may, dependent on the enzyme(s)applied, preferably be from about pH 5 to about pH 11, more preferablyfrom about pH 7 to about pH 11. The reaction time may preferably be inthe range of from about 15 minutes to about 3 hours.

In a preferred embodiment, the method of the invention is carried out inthe presence of other enzymes, in particular, a proteolytic enzyme, alipolytic enzyme, a cellulytic enzyme, an amylolytic enzyme, an oxidaseenzyme, or a peroxidase enzyme, or mixtures hereof.

The bio-polishing treatment according to the invention comprisestreating polyester or polyester containing fabrics or garments with anenzyme selected from the group consisting of a terephtalic acid diethylester hydrolytic enzyme (ETE hydrolytic enzyme), an ethyleneglycoldibenzyl ester hydrolytic enzyme (BEB hydrolytic enzyme), andcombinations of the foregoing, wherein said method is carried outwithout the presence of a detergent.

The polyester fabrics or garments may in one embodiment be a polyesterfabric consists of 100% polyester or essentially 100% polyester. Inanother embodiment the polyester containing fabric or garment is apolyester blend including any of the fabrics or garments mentioned inthe “Polyester Fabrics or Garments” section above.

In a preferred embodiment the fabric or garment bio-polishing is carriedout in the presence of a cellulytic enzyme, such as an endoglucanase,especially a fungal endoglucanase. However, further enzymes may beadded. In an embodiment the fabric or garment is further treated with anenzyme selected from the group consisting of proteases, amylases, othercellulases, peroxidases, oxidases, and pectinases, lipases other thanETE or BEB hydrolyases, and combinations of any of the foregoing.

Detergent

In the context of this invention, a detergent is synonymous with asurfactant, e.g., a nonionic surfactant, an anionic surfactant, acationic surfactant, an ampholytic surfactant, a zwitterionicsurfactant, and a semi-polar surfactant, or a mixture hereof.

Other Enzymes

The polyester hydrolytic enzyme of the invention may be added incombination with other enzyme(s).

Such enzymes include other proteases, lipases, amylases, cellulases,peroxidases and oxidases.

Proteases

Any protease suitable for use in alkaline solutions can be used.Suitable proteases include those of animal, vegetable or microbialorigin. Microbial origin is preferred. Chemically or geneticallymodified mutants are included. The protease may be a serine protease,preferably an alkaline microbial protease or a trypsin-like protease.Examples of alkaline proteases are subtilisins, especially those derivedfrom Bacillus, e.g., subtilisin Novo, subtilisin Carlsberg, subtilisin309, subtilisin 147 and subtilisin 168 (described in WO 89/06279).Examples of trypsin-like proteases are trypsin (e.g., of porcine orbovine origin) and the Fusarium protease described in WO 89/06270.

Preferred commercially available protease enzymes include those soldunder the trade names ALCALASE™, SAVINASE™, PRIMASE™, DURAZYM™, andESPERASE™ by Novozymes A/S (Denmark), those sold under the tradenameMAXATASE™, MAXACAL™, MAXAPEM™, PROPARASE™, PURAFECT™ and PURAPECT™ OXPby Genencor International Inc., (USA), and those sold under thetradename OPTICLEAN™ and OPTIMASE™ by Solvay Enzymes. Protease enzymesmay be incorporated into the compositions in accordance with theinvention at a level of from 0.00001% to 2% of enzyme protein by weightof the composition, preferably at a level of from 0.0001% to 1% ofenzyme protein by weight of the composition, more preferably at a levelof from 0.001% to 0.5% of enzyme protein by weight of the composition,even more preferably at a level of from 0.01% to 0.2% of enzyme proteinby weight of the composition.

Lipases

Any lipase suitable for use in alkaline solutions can be used. Suitablelipases include those of bacterial or fungal origin. Chemically orgenetically modified mutants are included.

Examples of useful lipases include a Humicola lanuginosa lipase, e.g.,as described in EP 258 068 and EP 305 216, a Rhizomucor miehei lipase,e.g., as described in EP 238 023, a Candida lipase, such as a C.antarctica lipase, e.g., the C. antarctica lipase A or B described in EP214 761, a Pseudomonas lipase such as a P. alcaligenes and P.pseudoalcaligenes lipase, e.g., as described in EP 218 272, a P. cepacialipase, e.g., as described in EP 331 376, a P. stutzeri lipase, e.g., asdisclosed in GB 1,372,034, a P. fluorescens lipase, a Bacillus lipase,e.g., a B. subtilis lipase (Dartois et al., (1993), Biochemica etBiophysica acta 1131, 253-260), a B. stearothermophilus lipase (JP64/744992) and a B. pumilus lipase (WO 91/16422).

Furthermore, a number of cloned lipases may be useful, including thePenicillium camembertii lipase described by Yamaguchi et al., (1991),Gene 103, 61-67), the Geotricum candidum lipase (Schimada, Y. et al.,(1989), J. Biochem., 106, 383-388), and various Rhizopus lipases such asa R. delemar_lipase (Hass, M. J et al., (1991), Gene 109, 117-113), a R.niveus lipase (Kugimiya et al., (1992), Biosci. Biotech. Biochem.56,716-719) and an R. oryzae lipase.

Especially suitable lipases are lipases such as M1 LIPASE™, LUMA FAST™and LIPOMAX™ (Genencor International Inc, USA), LIPOLASE™ and LIPOLASEULTRA™ (Novozymes A/S, Denmark), and LIPASE P “Amano” (AmanoPharmaceutical Co. Ltd.).

The lipases are normally incorporated in the composition at a level offrom 0.00001% to 2% of enzyme protein by weight of the composition,preferably at a level of from 0.0001% to 1% of enzyme protein by weightof the composition, more preferably at a level of from 0.001% to 0.5% ofenzyme protein by weight of the composition, even more preferably at alevel of from 0.01% to 0.2% of enzyme protein by weight of thecomposition.

Amylases

Any amylase (alpha and/or beta) suitable for use in alkaline solutionscan be used. Suitable amylases include those of bacterial or fungalorigin. Chemically or genetically modified mutants are included.Amylases include, for example, alpha-amylases obtained from a specialstrain of B. licheniformis, described in more detail in GB 1,296,839.Commercially available amylases are DURAMYL™, TERMAMYL™, FUNGAMYL™ andBAN™ (available from Novozymes A/S, Denmark) and RAPIDASE™ and MAXAMYLP™ (available from Genencor International Inc., USA).

The amylases are normally incorporated in the composition at a level offrom 0.00001% to 2% of enzyme protein by weight of the composition,preferably at a level of from 0.0001% to 1% of enzyme protein by weightof the composition, more preferably at a level of from 0.001% to 0.5% ofenzyme protein by weight of the composition, even more preferably at alevel of from 0.01% to 0.2% of enzyme protein by weight of thecomposition.

Cellulases

In the present context, the term “cellulase or “cellulolytic enzyme”refers to an enzyme, which catalyses the degradation of cellulose toglucose, cellobiose, triose and other cellooligosaccharides. Celluloseis a polymer of glucose linked by beta-1,4-glucosidic bonds. Cellulosechains form numerous intra- and intermolecular hydrogen bonds, whichresult in the formation of insoluble cellulose microfibrils. Microbialhydrolysis of cellulose to glucose involves the following three majorclasses of cellulases: endo-1,4-beta-glucanases (EC 3.2.1.4), whichcleave beta-1,4-glucosidic links randomly throughout cellulosemolecules; cellobiohydrolases (EC 3.2.1.91) (exoglucanases), whichdigest cellulose from the nonreducing end; and beta-glucosidases (EC3.2.1.21), which hydrolyse cellobiose and low-molecular-masscellodextrins to release glucose. Most cellulases consist of acellulose-binding domain (CBD) and a catalytic domain (CAD) separated bya linker rich in proline and hydroxy amino acid residues. In thespecification and claims, the term “endoglucanase” is intended to denoteenzymes with cellulolytic activity, especially endo-1,4-beta-glucanaseactivity, which are classified in EC 3.2.1.4 according to the EnzymeNomenclature (1992) and are capable of catalyzing (endo)hydrolysis of1,4-beta-D-glucosidic linkages in cellulose, lichenin and cerealbeta-D-glucans including 1,4-linkages in beta-D-glucans also containing1,3-linkages. Any cellulase suitable for use in alkaline solutions canbe used. Suitable cellulases include those of bacterial or fungalorigin. Chemically or genetically modified mutants are included.Suitable cellulases are disclosed in U.S. Pat. No. 4,435,307, whichdiscloses fungal cellulases produced from Humicola insolens. Especiallysuitable cellulases are the cellulases having color care benefits.Examples of such cellulases are cellulases described in European patentapplication No. 0 495 257, WO 91/17243 and WO 96/29397.

Commercially available cellulases include CELLUZYME™ and DENIMAX™produced by a strain of Humicola insolens, (Novozymes A/S), andKAC-500(B)™ (Kao Corporation).

Cellulases are normally incorporated in the composition at a level offrom 0.00001% to 2% of enzyme protein by weight of the composition,preferably at a level of from 0.0001% to 1% of enzyme protein by weightof the composition, more preferably at a level of from 0.001% to 0.5% ofenzyme protein by weight of the composition, even more preferably at alevel of from 0.01% to 0.2% of enzyme protein by weight of thecomposition.

In an embodiment of the method of the invention the cellulase may beused in a concentration in the range from 0.001-10 mg enzyme protein/mlsolution, preferably 0.005-0.3 mg enzyme protein/ml solution, especially0.001-0.003 mg enzyme protein/ml solution.

Peroxidases/Oxidases

Peroxidase enzymes are used in combination with hydrogen peroxide or asource thereof (e.g., a percarbonate, perborate or persulfate). Oxidaseenzymes are used in combination with oxygen. Both types of enzymes areused for “solution bleaching”, i.e. to prevent transfer of a textile dyefrom a dyed fabric to another fabric when said fabrics are washedtogether in a wash liquor, preferably together with an enhancing agentas described in e.g. WO 94/12621 and WO 95/01426. Suitableperoxidases/oxidases include those of plant, bacterial or fungal origin.Chemically or genetically modified mutants are included.

Peroxidase and/or oxidase enzymes are normally incorporated in thecomposition at a level of from 0.00001% to 2% of enzyme protein byweight of the composition, preferably at a level of from 0.0001% to 1%of enzyme protein by weight of the composition, more preferably at alevel of from 0.001% to 0.5% of enzyme protein by weight of thecomposition, even more preferably at a level of from 0.01% to 0.2% ofenzyme protein by weight of the composition.

Mixtures of the above mentioned enzymes are encompassed herein, inparticular a mixture of a protease, an amylase, a lipase and/or acellulase.

The enzyme, or any other enzyme incorporated in the composition, isnormally incorporated in the composition at a level from 0.00001% to 2%of enzyme protein by weight of the composition, preferably at a levelfrom 0.0001% to 1% of enzyme protein by weight of the composition, morepreferably at a level from 0.001% to 0.5% of enzyme protein by weight ofthe composition, even more preferably at a level from 0.01% to 0.2% ofenzyme protein by weight of the composition.

Materials and Methods

Enzymes:

Cutinase variant A is derived from Humicola insolens DSM 1800 and isdisclosed in WO 01/92502 and includes the following substitutions: E6Q,A14P, E47K, R51P, E179Q, GBD, N15D, S48E, A88H, N91H, A130V, R189V,T29M, T1661, L167P.

Cutinase variant B is derived from Humicola insolens DSM 1800 and isdisclosed in WO 01/92502 and includes the following substitutions: E6Q,A14P, E47K, R51P, E179Q, G8D, N15D, S48E, A88H, N91H, A130V, R189V.

Cellulytic enzyme: 43 kd endoglucanase (EG V) derived from Humicolainsolens DSM 1800, disclosed in WO 91/17243 as SEQ ID NO: 2.

Fabric

100% Dacron type 54 woven fabric and a 50%/50% polyester cotton singleknit were obtained from TestFabrics Inc. The 100% polyester fabric wasscoured, rinsed and dried as a preparation to enzymatic treatment. Theblended fabric was used as received. Experimental swatches were cut to14×14 cm. All swatches were conditioned overnight at a constanttemperature and humidity (21±20° C., 65±2% RH (Relative Humidity). Theweight was then measured and recorded.

Methods:

Bio-Polishing

Bio-polishing was carried out in a Launder-O-Meter (LOM) LP2 from AtlasElectric Devices Company. 20 Steel balls, buffer and enzyme were addedin 500 mL steel beakers. 50 mM sodium bicarbonate buffer was adjusted topH 8 and used for all experiments. The liquor ratio was 20:1 and theenzymes were dosed as U/ml (LU or ECU per liquor volume). Two swatcheswith a total weight of 7.0±0.1 g were used in each beaker and eachtreatment was run in duplicate. The beakers were loaded into a preheatedLaunder-O-Meter at 70° C. and incubated at 42 rpm for a specified time.

Following enzyme treatment, the residual enzyme activity on the swatcheswas inactivated at 80° C. in 2 g/L soda ash solution for 10 minutes.After inactivation, the swatches were rinsed and centrifuged in aconventional home washer and tumble dried for 60 minutes. All swatcheswere conditioned overnight at a constant temperature and humidity (21°C.±2° C. 65±2% RH).

Weight Loss

Final swatch weight was measured and recorded after conditioning todetermine weight loss. A mean weight loss was calculated by averagingthe weight loss data for all swatches in one treatment type.

Pilling

Pilling was measured according to ASTM D3786-87 (Pilling Resistance andOther Related Surface Changes of Textile Fabrics-Martindale PressureTester Method). A swatch from each beaker was tested and evaluated after125, 500 and 2000 revolutions on a Nu-Martindale Abrasion and PillingTester from James H. Heal & Co. Ltd. A pilling note was obtained byvisually comparing to a standard on a scale from 1-5, where 5 is nopilling and 1 is severely pilled. A mean pilling note was calculated byaveraging all swatches treated under the same conditions.

HPLC Analysis

Liquor samples were taken from each beaker after Launder-O-Meterincubation for HPLC analysis. The samples were filtered and loaded intovials. The samples were then injected on an Agilent 1100 series HPLC anddetected with a variable wavelength detector at 254 nm at 25° C. Themobile phase was a combination of solvent A—filtered deionized waterplus 0.1% trifluoroacetic acid and solvent B—100% acetonitrile. Agradient was used where solvent B increased from initial concentrationof 10% up to 95% at the end of the 19 minute run time. An Adsorbosil C₁₈column from Alltech (USA) was used. The flow rate was 0.8 ml/minute.After each injection the needle was rinsed in dimethylformamide and thecolumn was allowed to equilibrate for 5 minutes at initial conditions.Peak area counts for known degradation products of polyester wereaveraged for samples treated under the same conditions.

Lipase Activity (LU)

The esterase activity was measured according to Novozymes analyticalmethod 2001-07992-01 hereby incorporated by reference and available fromNovozymes A/S, Denmark, on request. In this assay, glycerol tributyratewas incubated with the esterase composition in 0.1 mM glycine buffer (pH7) at 30° C. over time. The lipase unit, or LU, is the amount of enzymewhich releases 1 micro mol of titratable butyric acid per minute. Theincubation is held for a minimum of two minutes and the resultingactivity is calculated and expressed in LU/g.

Cellulase Activity (ECU)

The cellulase activity was measured according to Novozymes analyticalmethod 302.02/01 hereby incorporated by reference and available fromNovozymes A/S, Denmark, on request. In this assay,carboxymethylcellulose (CMC) was incubated with the cellulasecomposition in 0.1 M phosphate buffer (pH 7.5) at 40° C. for 30 minutes.The reduction in viscosity was determined by a vibration viscometer andthe result was compared to a standard cellulase and expressed inendocellulase units as ECU/g.

Apparatus

Improvements in pilling resistance may be determined using theMartindale pilling tester (Swiss standard SN 198525) hereby incorporatedby reference).

EXAMPLES

The invention is further illustrated with reference to the followingexamples which are not intended to be in any way limiting to the scopeof the invention as claimed.

Example 1 Hydrolytic Activity

This example described an assay for determining the terephtalic aciddiethyl ester (ETE) and/or an ethyleneglycol dibenzyl ester (BEB)hydrolytic activity of an enzyme.

ETE Hydrolytic Activity

An ETE hydrolytic enzyme of the invention is an enzyme capable ofhydrolyzing terephtalic acid diethyl ester (ETE), as determined by thefollowing assay.

In a test tube, 0.250 ml of 0.20 M glycylglycine pH 8.5, and 0.250 ml of10.0 mM terephthalic acid diethyl ester (ETE) in 1,4-dioxane, is addedto 2.000 ml of de-ionized water.

The mixture is pre-incubated under stirring at 30° C. for approximately15 minutes, followed by the addition of 25.0 micro g of enzyme in thelowest possible volume. This mixture is then subjected to incubationunder stirring at 30° C. for 16 hours.

The reaction mixture is analyzed on a reverse phase HPLC, ODS (octadodecyl silicate) column, and eluted with increasing concentration ofacetonitrile and decreasing concentration of 200 mM NaPO₄, pH 3.0.

Detection of the reaction products is carried out spectrophotometricallyat 240 nm, at which wavelength terephtalic acid and terephtalatederivatives adsorb.

BEB Hydrolytic Activity

A BEB hydrolytic enzyme of the invention is an enzyme capable ofhydrolyzing ethyleneglycol dibenzyl ester (BEB). Dependent on the amountof dioxane present in the assay (BEB is only partially dissolved in a10% dioxane solution, but fully dissolved in a 30% dioxane solution),the BEB hydrolytic enzyme of the invention may be a BEB¹⁰ hydrolyticenzyme or a BEB³⁰ hydrolytic enzyme, as determined by the followingassays.

BEB¹⁰ Hydrolytic Activity

In a test tube, 0.250 ml of 0.20 M glycylglycine pH 8.5, and 0.250 ml of10.0 mM ethyleneglycol dibenzyl ester (BEB) in 1,4-dioxane, is added to2.000 ml of de-ionized water.

The mixture is pre-incubated under stirring at 30° C. for approximately15 minutes, followed by the addition of 25.0 micro g of enzyme in thelowest possible volume. This mixture is then subjected to incubationunder stirring at 30° C. for 16 hours.

The reaction mixture is analyzed on a reverse phase HPLC, ODS (octadodecyl silicate) column, and eluted with increasing concentration ofacetonitrile and decreasing concentration of 200 mM NaPO₄, pH 3.0.

Detection of the reaction products is carried out spectrophotometricallyat 240 nm, at which wavelength terephtalic acid and terephtalatederivatives adsorb.

BEB³⁰ Hydrolytic Activity

In a test tube, 0.250 ml of 0.20 M glycylglycine pH 8.5, 0.250 ml of10.0 mM ethyleneglycol dibenzyl ester (BEB) in 1,4-dioxane, and 0.500 mlof 1,4-dioxane, is added to 1.500 ml of de-ionized water. The mixture ispre-incubated under stirring at 30° C. for approximately 15 minutes,followed by the addition of 25.0 micro g of enzyme in the lowestpossible volume. This mixture is then subjected to incubation understirring at 30° C. for 16 hours.

The reaction mixture is analyzed on a reverse phase HPLC, ODS (octadodecyl silicate) column, and eluted with increasing concentration ofacetonitrile and decreasing concentration of 200 mM NaPO₄, pH 3.0.

Detection of the reaction products is carried out spectrophotometricallyat 240 nm, at which wavelength terephtalic acid and terephtalatederivatives adsorb.

Microbial Sources

A number of enzymes from different microbial sources were subjected tothe assay for determining BEB and ETE hydrolytic activity, and theresults obtained are presented in Table 1, below: TABLE 1 Enzymes havingBEB and/or ETE Hydrolytic Activity; % Degradation of Substrate. EnzymeSubstrate Microbial Source BEB³⁰ BEB¹⁰ ETE Humicola insolens ¹⁾ 100 10095 Candida antarctica ²⁾ 60 100 100 Pseudomonas cepacia ³⁾ 95 — 60Candida cylindracea ⁴⁾ 0 100 15 GA⁵⁾ 0  0 5¹⁾ Humicola insolens cutinase (actually a lipase also having cutinaseactivity) obtained from the strain DSM 1800 as described in Example 2 ofU.S. Pat. No. 4,810,414.²⁾ Candida antarctica Component B obtained as described in Example 10 ofWO 88/02775.³⁾ Pseudomonas cepacia obtained as described in EP 331,376.⁴⁾ Candida cylindracea (syn. Candida rugosa) lipase obtained from NipponOil & Fats Co. Ltd., Japan).⁵⁾Glucosaminated LIPOLASE ™ obtained as described in Example 7A of WO95/09909.

Example 2 Reduced Pilling Propensities

140 ml 50 mM Sodium Bicarbonate buffer pH 8 was added to eachLaunder-O-Meter beaker including 20 steel balls. The beakers wereequilibrated at 70° C. Two woven 100% polyester swatches (approx. 14×14cm each) were added to each Launder-O-Meter beaker. Enzyme (Humicolainsolens cutinase) was added at different dosages and with and withoutthe addition of Triton X-100 dosed at 1 g/L including a blank withoutenzyme. The swatches were incubated for 4 hours at 70° C. in theLaunder-O-Meter turning at 42 rpm. After incubation the swatches weregiven a short rinse in a household laundry machine and tumble dried for1 hour.

Pilling measurements are measured using the Martindale pilling tester(Swiss standard SN 198525).

The results are shown in table 2. TABLE 2 Pilling note for 100%polyester fabric Cutinase Revolutions Blank - Cutinase 50 LU/mL and 1g/L Triton tested no enzyme 50 LU/mL 1 g/L Triton X-100 X-100 only 125rev. 2.75 4 3.25 3.5 500 rev. 2 3.25 2.5 2.75 2000 rev.  1.5 3.25 2 2.5A pilling note is obtained by visually comparing to a standard on ascale from 1-5, where 5 is no pilling and 1 is severely pilled. A meanpilling note is calculated by averaging all swatches treated under thesame conditions.

Example 3 Bio-Polishing of 100% Polyester

The bio-polishing effect of two Humicola insolens ETE and/or BEBhydrolytic enzymes (in the form of two cutinase variants, referred to ascutinase variant A and B, respectively), were tested on 100% polyesteras described in the “Materials & Methods” section. The 100% polyesterfabric was scoured, rinsed and dried as a preparation to enzymatictreatment. Initially the polyester fabric was enzymatically degradedwith cutinase variant A and cutinase variant B, respectively. Cutinasevariant B gave higher weight loss than that of cutinase variant A,whereas cutinase variant A gave little to no weight loss compared to ablank (see FIG. 1). A blank is defined as a treatment performed where noenzyme is added.

Both enzymes gave improvement on pilling note compared to the blank at2000 revolutions (see FIG. 2). Although there was little to no weightloss for cutinase variant A, an improvement in pilling note was observedas compared to the blank.

Additionally, HPLC results were measured to detect enzymatic degradation(see FIG. 3). Cutinase variant B shows considerably higher degradationaccording to HPLC area count of degradation products of polyester. Thiscorrelates with higher pilling note and weight loss. However, FIG. 3does illustrate that both enzymes are acting on the polyester as asubstrate and the pilling prevention data is an enzymatic hydrolysiseffect.

The test demonstrates that 100% polyester fabric can be treated with ETEand/or BEB hydrolytic enzymes to impart a durable bio-polished finishwithout the presence of a detergent.

Example 4 Bio-Polishing of Polyester/Cotton Blends

The bio-polishing effect of an ETE and/or BEB hydrolytic enzyme(cutinase variant A) in combination with a 43 kD cellulase from Humicolainsolens, DSM 1800, was tested on a 50%/50% polyester cotton asdescribed in the “Materials & Methods” section. The blended fabric wasused untreated.

Two dose response trials were performed. The first dose response wasmaintaining a 0.75 ECU/ml dose of cellulase and increasing the dosage ofCutinase variant A from 0 to 50 LU/ml. The second dose response trialwas maintaining a 50 LU/ml dose of cutinase variant A and increasing thedosage of cellulase from 0 to 1 ECU/ml.

All data from polyester/cotton blend bio-polishing is shown is Table 3.TABLE 3 Bio-Polishing of 50%/50% Polyester/Cotton Blend, LOM treatment,2 hours, pH 8 at 70° C. Enzyme Treatment Evaluation Dose Cutinase HPLCarea Dose Cellulase variant A Weight Pilling at count at (ECU/ml)(LU/ml) Loss % 2000 rev. 254 nm 0 0 1.0 1.25 9 0.75 0 1.1 2.5 8 0.75 101.3 3 1888 0.75 20 1.4 2.3 2713 0.75 30 1.4 3 3058 0.75 50 1.6 3 3503 050 1.3 1.5 3699 0.25 50 1.6 2.8 3616 0.5 50 1.6 2.5 3524 0.75 50 1.6 33530 1 50 1.7 3 3559

The weight loss measured for each dose response increased with increaseddosage of both enzymes.

Both cellulase and Cutinase variant A gave an improvement in pillingnote as compared to the blank. When both enzymes are combined, the mostsignificant improvement in pilling note is observed at almost all dosesevaluated.

HPLC results were measured to detect enzymatic degradation of thepolyester polymer due to cutinase activity. The area count ofdegradation products of polyester increases as the enzyme doseincreases. As the cellulase dose increases, the area count does notchange because the dosage of Cutinase variant A was constant.

The test demonstrates that a polyester cotton blend fabric can betreated with a cellulase combined with a ETE and/or BEB hydrolyticenzyme to impart a durable bio-polished finish without the presence of adetergent.

1. A method of reducing the pilling propensity of a polyester fabric orgarment, which comprises treating the fabric or garment with an enzymeselected from the group consisting of a terephtalic acid diethyl esterhydrolytic enzyme (ETE hydrolytic enzyme), an ethyleneglycol dibenzylester hydrolytic enzyme (BEB hydrolytic enzyme), and combinations of theforegoing, wherein said method is carried out without the presence of adetergent.
 2. The method according to claim 1, wherein the fabric orgarment is treated with a terephtalic acid diethyl ester hydrolyticenzyme (ETE hydrolytic enzyme).
 3. The method according to claim 1,wherein the fabric or garment is treated with an ethyleneglycol dibenzylester hydrolytic enzyme (BEB hydrolytic enzyme).
 4. The method accordingto claim 1, further comprising treating said fabric or garment with anenzyme selected from the group consisting of proteases, amylases,cellulases, peroxidases, oxidases, pectinases, lipases other than ETE orBEB hydrolyases, and combinations of any of the foregoing.
 5. The methodaccording to claim 4, wherein said fabric or garment is treated with acellulytic enzyme.
 6. A method of color clarification of a polyesterfabric or garment, which comprises treating the fabric or garment withan enzyme selected from the group consisting of a terephtalic aciddiethyl ester hydrolytic enzyme (ETE hydrolytic enzyme), anethyleneglycol dibenzyl ester hydrolytic enzyme (BEB hydrolytic enzyme),and combinations of the foregoing, wherein said method is carried outwithout the presence of a detergent.
 7. The method according to claim 6,wherein the fabric or garment is treated with a terephtalic acid diethylester hydrolytic enzyme (ETE hydrolytic enzyme).
 8. The method accordingto claim 6, wherein the fabric or garment is treated with anethyleneglycol dibenzyl ester hydrolytic enzyme (BEB hydrolytic enzyme).9. The method according to claim 6, further comprising treating saidfabric or garment with an enzyme selected from the group consisting ofproteases, amylases, cellulases, peroxidases, oxidases, and pectinases,lipases other than ETE or BEB hydrolyases, and combinations of any ofthe foregoing.
 10. The method according to claim 6, wherein said fabricor garment is treated with a cellulytic enzyme.
 11. A method ofbio-polishing polyester containing fabrics or garments, which methodcomprises treating said fabric or garment with an enzyme selected fromthe group consisting of a terephtalic acid diethyl ester hydrolyticenzyme (ETE hydrolytic enzyme), an ethyleneglycol dibenzyl esterhydrolytic enzyme (BEB hydrolytic enzyme), and combinations of theforegoing, wherein said method is carried out without the presence of adetergent.
 12. The method of claim 11, wherein the polyester containingfabric or garment consists of essentially 100% polyester.
 13. The methodof claim 11, wherein the polyester fabric or garment is a polyesterblend, such as a polyester and cellulosic blend, including polyester andcotton blends; a polyester and wool blend; a polyester and silk blend; apolyester and acrylic blend; a polyester and nylon blend; a polyester,nylon and polyurethane blend; a polyester and polyurethane blend, rayon(viscose), cellulose acetate and tencel.
 14. The method according toclaim 11, wherein said fabric or garment is further treated with acellulytic enzyme.
 15. The method according to claim 11, wherein thefabric or garment is treated with a terephtalic acid diethyl esterhydrolytic enzyme (ETE hydrolytic enzyme).
 16. The method according toclaim 11, wherein the fabric or garment is treated with anethyleneglycol dibenzyl ester hydrolytic enzyme (BEB hydrolytic enzyme).17. The method according to claim 11, further comprising treating saidfabric or garment with an enzyme selected from the group consisting ofproteases, amylases, other cellulases, peroxidases, oxidases, andpectinases, lipases other than ETE or BEB hydrolyases, and combinationsof any of the foregoing.